Pancreatic adenocarcinoma (PDA) has incredibly poor patient prognosis with a 5- year survival rate less than 5% with most patients succumbing to cancer metastasis. PDA is predicted to become the second leading cause of cancer-related deaths in the United States by the year 2020. The molecular genetic alterations associated with the metastatic cascade - invasion away from primary tumor, entry into the bloodstream, and extravasation and colonization at a distant site - remain unclear and understudied. Understanding this process is imperative for developing more effective therapies. The goal of this proposal is to use an in vivo platform to identify specific genes and pathways that drive metastasis (so-called metastasis virulence genes) in PDA. To achieve this goal, we will use genetically engineered mouse models of PDA and deep- sequencing technology to identify and validate novel exome and transcriptome alterations that associate with metastatic cascade. In Aim 1, exome and mRNA sequencing of matched primary tumor and metastasis from PDA mouse models will be performed to identify novel variants. Potential variants will be compared to available human PDA datasets to identify similar mutations. We will functionally validate that these alterations can mediate metastatic disease in Aim 2. Targeting (using RNAi and ORF libraries) will be used to specifically delete or induce the expression of gene(s) of interest identified in Aim 1. Validation of mutations of interest will be done via orthotopic transplantatio of targeted pancreatic cancer cell lines. In sum, this proposal will identify the exact genomic and transcriptomic landscape driving metastasis in mouse models of PDA and will provide critical information for the development of targeted therapeutics.